QQ客服 官方微博 反馈留言

	E-cadherin前体蛋白抗体的制备与鉴定; Preparation and Identification of the Antibody Against the Precursor Protein of E-cadherin
	王继峰 ; 叶志云 ; 林舒勇
	2011-01
关键词	E-cadherin pre 抗体 pGST E-cadherin pre antibody
英文摘要	通信作者: lin sy@ xmu. edu. cn; [中文文摘]E-cadherin是一种重要的肿瘤转移抑制因子,它的功能异常能够导致肿瘤细胞的转移.细胞内成熟的E-cadherin由其前体蛋白剪切而成,目前还没有针对该前体蛋白的特异性抗体.为了得到能用于检测内源性E-cadherin前体蛋白(E-cadherin pre)的抗体,对E-cadherin基因用限制性内切酶PstⅠ和PvuⅡ进行双酶切,获取E-cadherin基因的N-端片段,将其重组入原核表达载体pGEX-4T-2中,在大肠杆菌BL21(DE3)菌株中表达重组蛋白,经谷胱甘肽-琼脂糖珠子纯化后,得到较高纯度的E-cadherin前体抗原.用该抗原免疫新西兰兔,获得抗血清,进一步纯化血清得到抗E-cadherinpre的多克隆抗体.用Western blot检测了外源性和内源性E-cadherin pre,发现该抗体效价高且特异性强;免疫荧光实验表明该抗体可以用于细胞染色,为深入研究E-cadherin pre在细胞内的功能奠定了基础. [英文文摘]E-cadherin, an important tumor suppressor which was defected highly correlates with tumor metastasis was derived from its precursor protein. As for the commercial unavailability , the precursor form o f E-cadherin can not be detected at endo genous levels.In order to get qualified antibody agianst E-cadher in pr e, Ps t I-Pv u II fragment (amino acids 31 to 164) N-terminal in the precursor region of E-cadher in was ligated to pGST-parallel vector, expressed in the E. coli. strain BL-21. The fusion protein was purified by GST beads, and rabbits were immunized. 9 weeks later , the sera of the rabbits were har vested and an anti-Pre-E-cadherin polyclonal antibody was purified. The precursor of E-cadher in was successfully detected with great sensit ivity and specificity by western blotting using t his anti- Pre-E-cadher in antibody in the lysate of 293T cells transfected with pCMV5-c-myc E-cadherin. Fur thermore, the antibody lays a good foundation for fur ther research on the function of E-cadherin pre, especially on its role in signal transduction pathways.; 福建省自然科学基金资助项目(2008J0109)
语种	中文
出版者	《厦门大学学报(自然科学版) 》编辑部
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/14994]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	王继峰,叶志云,林舒勇. E-cadherin前体蛋白抗体的制备与鉴定, Preparation and Identification of the Antibody Against the Precursor Protein of E-cadherin[J],2011.
APA	王继峰,叶志云,&林舒勇.(2011).E-cadherin前体蛋白抗体的制备与鉴定..
MLA	王继峰,et al."E-cadherin前体蛋白抗体的制备与鉴定".(2011).

个性服务

查看访问统计

相关权益政策

暂无数据

收藏/分享

除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。